枳椇果梗多糖调节谷氨酸代谢和紧密连接蛋白表达改善酒精对小鼠神经行为影响  

Hovenia dulcis Fruit Peduncle Polysaccharides Modulate Glutamate Metabolism and Tight Junction Protein Expressions to Attenuate the Neurotoxic Effects of Alcohol on Mice

作　　者：张玉超 袁子飞 刘良禹 朱思洁 杨永 郑代梅 刘旭东 ZHANG Yuchao;YUAN Zifei;LIU Liangyu;ZHU Sijie;YANG Yong;ZHENG Daimei;LIU Xudong(Department of Brewing Engineering,Moutai Institute,Renhuai 564507,China;Department of Food Science and Engineering,Moutai Institute,Renhuai 564507,China;College of Food Science and Engineering,Ocean University of China,Qingdao 266071,China)

机构地区：[1]茅台学院酿酒工程系,贵州仁怀564507 [2]茅台学院食品科学与工程系,贵州仁怀564507 [3]中国海洋大学食品科学与工程学院,山东青岛266071

出　　处：《食品工业科技》2025年第9期372-380,共9页Science and Technology of Food Industry

基　　金：贵州省基础研究(自然科学)计划项目(黔科合基础-ZK[2022]一般541,黔科合基础-ZK[2022]一般542);遵义市科技计划项目(遵市科合HZ字(2024)387号);茅台学院酒产业研究院2024年专项课题(MTXYJCY009);茅台学院高层次人才科研启动经费项目(mygccrc[2022]010);贵州省大学生创新创业训练计划项目(S202214625019,S202214625021)。

摘　　要：目的:本研究旨在明确枳椇果梗多糖(HDPs)对酒精暴露所致的小鼠神经行为异常的改善效果,并探究谷氨酸代谢和紧密连接蛋白表达在其中的作用。方法:雄性C57BL/6小鼠按114μL/20 g剂量连续酒精灌胃14 d,建立酒精暴露模型,同时设置干预组进行HDPs干预(114μL/20 g酒精+100 mg/kg HDPs)。应用行为学实验(旷场实验、高架十字迷宫实验)评估神经行为学变化,采用气相色谱法测定小鼠血液中乙醇浓度,γ-H2AX荧光检测小鼠脑海马组织DNA损伤,免疫组化分析检测小鼠脑组织中紧密连接蛋白Claudin-1和ZO-1的表达,并通过超高效液相色谱-四级杆飞行时间质谱法(UPLC-Q-TOF-MS)代谢组学技术对小鼠脑组织代谢物进行分析。结果:HDPs可有效降低酒精暴露小鼠血液乙醇浓度,由4.69±0.29 g/L降至1.64±0.104 g/L;改善酒精暴露所致的小鼠神经行为异常,旷场实验中,与酒精组相比,HDPs干预组总路程显著提升至27340±3304 cm(P<0.05),平均速度显著提升至67.4±13.4 cm/s(P<0.05),不动时间缩短29%(P<0.05);高架十字迷宫实验中,与酒精组相比,HDPs干预组闭臂停留时间显著减少至195.6±10.3 s(P<0.05),开放臂进入次数显著增加26%(P<0.05);还可降低酒精诱导的脑组织氧化应激与DNA损伤水平,ROS、MDA分别降低5.4%、29.5%(P<0.05),T-AOC提高10.9%,上调脑海马组织中Claudin-1(2.2倍)和ZO-1(10%)蛋白的表达;并调节脑组织谷氨酸代谢通路,提高甘氨酸(19.7%)、谷光甘肽(25%)、琥珀酸(22.6%)等代谢物水平。结论:HDPs可有效改善酒精对小鼠神经行为的影响,其机制或可能通过抗氧化、保护紧密连接蛋白和调节谷氨酸代谢通路发挥作用,研究结果可为扩展枳椇资源在食品领域中的应用提供理论依据。Objective:This study aimed to investigate the protective effects of Hovenia dulcis fruit peduncle polysaccharides(HDPs)on alcohol-induced neurobehavioral alterations in mice and to elucidate whether HDPs mitigate alcohol-induced neuronal damage by modulating glutamate metabolic pathway and the expression of tight junction protein.Methods:Male C57BL/6 mice were administered alcohol intragastrically at a dose of 114μL/20 g for 14 d to establish an alcohol exposure model,and an intervention group was set up for HDPs intervention(114μL/20 g alcohol+100 mg/kg HDPs).Behavioral experiments(open field test,elevated plus maze test)were used to assess changes in neurobehavior,gas chromatography was used to determine ethanol concentrations in mouse blood.γ-H2AX fluorescence was used to detect DNA damage in mouse hippocampal tissue,and immunohistochemical analysis was used to detect the expression of tight junction proteins Claudin-1 and ZO-1 in mouse brain tissue.Metabolites in mouse brain tissue were analyzed using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)metabolomics technology.Results:HDPs effectively reduced blood ethanol concentration in alcohol-exposed mice(from 4.69±0.29 g/L to 1.64±0.104 g/L)and ameliorated alcohol-induced neurobehavioral abnormalities.In the open field test,compared to the alcohol group,HDPs intervention significantly increased total distance traveled(27340±3304 cm,P<0.05)and average velocity(67.4±13.4 cm/s,P<0.05),while reduced immobility time by 29%(P<0.05).The elevated plus maze test revealed that HDPs treatment decreased closed-arm dwelling time(195.6±10.3 s,P<0.05)and increased open-arm entries by 26%(P<0.05)compared to the alcohol group.Additionally,HDPs alleviated alcohol-induced oxidative brain damage by reducing ROS levels(5.4%)and MDA content(29.5%)(P<0.05)respectively,while enhanced total antioxidant capacity(TAOC)by 10.9%.It upregulated hippocampal expression of tight junction proteins Claudin-1(2.2-fold)and ZO-1(10%)

关 键 词：枳椇果梗多糖(HDPs) 酒精 神经行为 谷氨酸代谢 紧密连接蛋白 

分 类 号：TS201[轻工技术与工程—食品科学]